Composition for the regulation of the human immune system and the prevention and treatment of diseases thereof

ABSTRACT

A nutritional supplement composed of phytosterols, anti-oxidants, and other complexes, including essential fatty acids, amino acids, peptides, proline rich polypeptides and digestive enzymes is described. The nutritional supplement may be used by individuals suffering from or at risk of developing immune system diseases; breast cancer, colon and prostate cancer; HIV infection; high cholesterol; or enlarged prostate. In the preferred form, the invention comprises phytosterols and antioxidants, together with essential fatty acids derived from amino acids, short chain peptides, proline rich polypeptides and digestive enzymes, and a microcellulose filler.

PRIOR APPLICATION INFORMATION

This application is a continuation-in-part of U.S. patent applicationSer. No. 11/106,446, filed Apr. 15, 2005, which is acontinuation-in-part of International Application PCT CA03/01451, filedSep. 24, 2003 which in turn claims the benefit of Canadian PatentApplication 2,407,429, filed Oct. 16, 2002.

FIELD OF THE INVENTION

This invention relates to the field of nutritional supplements.

BACKGROUND OF THE INVENTION

Phytosterols are fats that are present in all plants, including fruitsand vegetables. Although structurally similar to cholesterol, thesterols synthesized by animals and plants differ in the nature of theirside chain (see FIG. 1) (Allayee et al. 2000, Science 290: 1709-1711).

In animals, cholesterol is the most abundant sterol. In plants, morethan 40 sterols have been identified, of which beta-sitosterol,stigmasterol, and campesterol are the most common (Hicks and Moreau2001, Food Technol 55: 63-67). These phytosterols occur in free form orare esterified to free fatty acids, sugar moieties or phenolic acids(Baker et al. 1999, Food Chem Toxicol 37: 13-22). Studies havedemonstrated that beta-sitosterol possesses anti-inflammatory and immunemodulating properties. It has been estimated that consumption of 3 g/dayof phytosterols could reduce the risk of heart disease by 15% to 40%,depending on age and other dietary factors (Hicks and Moreau 2001).However, the western diet typically contains 100-300 mg of plant sterols(Nguyen 1999, J Nutr 129: 2109-2112).

Enzogenol™ is an example of a water soluble extract of monomeric andoligomeric proanthocyanidins, flavonoids, flavonoid glycosides, estersand natural organic acids prepared from the bark of Pinus radiate by apure water extraction process (Shand et al. 2003, Phytother Res 17:490-494). The extract has been shown to have in vitro antioxidant actionas measured by inhibition of micelle oxidation, red blood cell hemolysisand a nitro blue tetra zolium enzymatic method (Gieseg and Baird 1998,Free Rad Biol Med 25: S104; Wood et al. 2002, Food Chem 77: 155-161).

Omega 3, 6 and 9 fatty acids, such as found in Cellasate, have astructural role in phospholipids of all cell membranes in the body,influencing membrane viscosity and permeability (Drevon 1992, Nutr Rev50: 38-45). Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)are thought to be responsible for beneficial effects, such as preventionand management of coronary heart disease and hypertension (Simopoulos1999, Am J Clin Nutr 70: 560S-569S).

SUMMARY OF THE INVENTION

According to a first aspect of the invention, there is provided a methodof modifying blood lipid parameters comprising administering to anindividual in need of such treatment an effective amount of acomposition comprising:

10% to 90% phytosterols;

5% to 85% essential fatty acid complexes; and

5% to 85% antioxidants.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are now described. All publications mentioned hereunderare incorporated herein by reference.

As used herein, the term “treating” in its various grammatical formsrefers to curing, reversing, attenuating, alleviating, minimizing,suppressing or halting the deleterious effects of a disease state,disease progression, disease causitive agent or other abnormalcondition.

Described herein is a new composition or nutritional supplement composedof natural products. In some embodiments, the supplement may be used byindividuals desirous of regulating their immune system, for example, toprevent and treat diseases thereof. As discussed below, the compositioncombines phytosterols with anti-oxidants and fatty acid complexes.

The composition has also been demonstrated to be beneficial in themodification of blood lipid parameters, as discussed below.Specifically, the composition or supplement has been shown to reduce thetotal cholesterol (TC) to high density lipoprotein (HDL) ratio (TC/HDL)as well as to reduce the low density lipoprotein (LDL) to HDL ratio(LDL/HDL) compared to a placebo-treated control, as discussed below. Aswill be appreciated by one of skill in the art, the composition orsupplement at formulae and/or dosages described herein is in someembodiments of the invention administered to an individual in need ofsuch treatment or used for modifying the blood lipid parameters of saidindividual, for example, reducing the TC/HDL ratio or reducing theLDL/HDL ratio. As will be appreciated by one of skill in the art,examples of such individuals include but are by no means limited toindividuals having, at risk of having or suspected of havinghypercholesterolemia or otherwise having or suspected of having elevatedcholesterol levels or reduced HDL levels.

As discussed below, administration of the composition or supplement toan individual in need of such treatment reduced basophil and IL-6 levelsin said individual compared to a second individual administered aplacebo control.

In another embodiment, administration of the supplement as describedherein to an individual in need of such treatment reduced LDL levels,increased HDL levels, reduced the LDL/HDL ratio or reduced the totalcholesterol to HDL ratio in said individual compared to a secondindividual administered a placebo control.

In one embodiment, the composition or supplement comprises: 10% to 90%phytosterols, 5% to 85% essential fatty acid complexes and 5-85%antioxidants. In other embodiments, the supplement comprises 20% to 90%phytosterols, 5% to 75% essential fatty acid complexes and 5-75%antioxidants. In another embodiment, the composition or supplementcomprises: 10% to 80% phytosterols, 10% to 80% essential fatty acidcomplexes and 10-80% antioxidants. In other embodiments, the supplementcomprises 20% to 80% phytosterols, 10% to 80% essential fatty acidcomplexes and 10 to 80% antioxidants. In a preferred embodiment, thesupplement comprises 29% to 75% phytosterols; 10% to 50% essential fattyacid complexes; and 5% to 50% antioxidants. The supplement orcomposition may be arranged in the form of a capsule or other suitable,ingestible format, such as a tablet, pill or other similar form known inthe art.

In a preferred embodiment, the supplement or composition comprises 29%to 75% beta-sitosterol; 10% to 50% essential fatty acid complexes; and5% to 50% antioxidants.

In a yet further preferred embodiment, there is provided a single-dosesupplement or composition comprising: 300 mg phytosterols; 50 mgessential fatty acid complexes; 20 mg antioxidants; and 30 mgmicro-cellulose filler. As discussed herein, this composition may betaken once daily or twice daily.

In some embodiments, the above-described compositions are used fortreating, preventing or ameliorating symptoms associated with an immunesystem disease. In these embodiments, the composition or supplement at adosage, concentration or range described above is administered to anindividual in need of such treatment. Examples of immune system diseasesinclude but are by no means limited to colds, flu, diabetes, allergies,asthma, pneumonia, fibromyalgia, HIV infection, hepatitis C infection,and multiple sclerosis. In a preferred embodiment, the composition atdosages, concentrations or ranges described above is used to treat orprevent pneumonia, fibromyalgia or a hepatitis C infection. In a yetpreferred embodiment, the composition or supplement described above isused to treat an immune system disease as described above byadministering the composition at a concentration or dosage as describedto an individual in need of such treatment.

Accordingly, in yet another embodiment, there is provided a method ofdecreasing the LDL/HDL ratio or reducing the TC/HDL ratio in anindividual in need of such treatment comprising administering to saidindividual an effective amount of a supplement comprising 10% to 90%phytosterols, 5% to 85% essential fatty acid complexes and 5-85%antioxidants. In other embodiments, the supplement comprises 20% to 90%phytosterols, 5% to 75% essential fatty acid complexes and 5-75%antioxidants. In another embodiment, the composition or supplementcomprises: 10% to 80% phytosterols, 10% to 80% essential fatty acidcomplexes and 10-80% antioxidants. In other embodiments, the supplementcomprises 20% to 80% phytosterols, 10% to 80% essential fatty acidcomplexes and 10 to 80% antioxidants. In a preferred embodiment, thesupplement comprises 29% to 75% phytosterols; 10% to 50% essential fattyacid complexes; and 5% to 50% antioxidants. In a further preferredembodiment, the supplement or composition comprises 29% to 75%beta-sitosterol; 10% to 50% essential fatty acid complexes; and 5% to50% antioxidants. In a yet further preferred embodiment, there isprovided a single-dose supplement or composition comprising: 300 mgphytosterols; 50 mg essential fatty acid complexes; 20 mg antioxidants;and 30 mg micro-cellulose filler. As discussed herein, this compositionmay be taken once daily or twice daily.

The inherent ingenuity of the invention is the discovery of the range ofthe amount of beta-sitosterol that is the most effective to achieve thegoal of regulating the Human Immune System and the prevention and/ortreatment of diseases thereof. As a result, this new composition needonly be taken once a day to maintain general health.

The further uniqueness of the invention is the discovery that combiningphytosterols with anti-oxidants and fatty acid complexes produces asynergistic effect whereby the cumulative total effect of thecomposition is greater than the sum of the individual effects of therespective components. As a result, this new composition optimizes theimmune system to deal with viral and bacterial infections, while alsoprotecting the body from degeneration due to free radical damage.

Additionally, the invention is original and novel because it can notonly assist in preventing possible causes of cancer, but, it inhibits orreduces cancer cell growth in breast, colon and prostate cancer cases,and facilitates the removal of the dead tissue arising from the death ofthe said tumors. That is, the composition can modulate or optimize, asopposed to only boosting, the immune system, as discussed below.

The utility of said invention is further demonstrated and confirmed bythe fact that it accomplishes its goals with none of the physical orpsychological side effects of traditional treatments such as radiation,chemotherapy and surgery.

The basis of the present invention is the use of a combination ofphytosterols, anti-oxidants and fatty acid complexes for the followingpurposes:

1. To modulate the Human Immune System;

2. To prevent or treat diseases of the Human Immune System, for examplebut by no means limited to:

Common cold and flu;

Microorganism infections, for example viral, yeast and bacterialinfections;

Allergies;

Rheumatoid Arthritis;

Lupus;

Chronic Fatigue Syndrome;

Fibromyalgia;

Asthma;

Diabetes; and

Eczema.

3. To inhibit or reduce cancer cell growth in breast, colon and prostatecancer cases;

4. To maintain CD4 Lymphocyte counts in HIV patients;

5. To treat enlarged prostate problems;

6. As an anti-inflammatory agent; and

7. Modifying blood lipid parameters, as discussed below.

As discussed above, the compositions described herein act to modulatethe immune system. Specifically, the compositions act to restore normalfunctions in an individual in need thereof. As will be apparent to oneof skill in the art, this explains how the compositions are able totreat diseases or conditions characterized by immune activation, such asautoimmune disorders such as arthritis and inflammation, as well asdiseases characterized by immune suppression or deficiency, such asacquired immune deficiency syndrome. While not wishing to be bound to aspecific theory, the inventors believe that the composition acts tomodulate immune function such that an immune system that is actingabnormally, that is, is either depressed or chronically activated,returns to normal functioning, that is, returns to the state ofactivation found in most individuals who are not currently undergoing achallenge to their immune system.

Although preferred compositions to achieve these ends are listed below,they are by no means to be interpreted as limiting the broad scope ofthe present invention as various other combinations of phytosterols,anti-oxidants and fatty acid complexes may achieve the same beneficialresults.

The preferred composition of the present application consists of 400 mgcapsules containing 300 mg of phytosterols (a minimum of which is 117 mgof beta-sitosterol), 50 mg essential fatty acid complexes, 20 mg broadspectrum antioxidants, and 30 mg micro-cellulose filler. As will beappreciated by one of skill in the art, this represents an example of aneffective amount of the composition. Other suitable concentrations anddosages may be determined using the ranges described herein. As will beapparent to one of skill in the art, as used herein, “effective amount”refers to an amount that achieves the desired effect.

In another preferred form, the composition comprises from 29% to 75%beta-sitosterol, 10% to 50% essential fatty acid complexes and from 5%to 50% antioxidants. As will be appreciated by one of skill in the art,other suitable, pharmaceutically acceptable fillers may also be used inthe invention.

In other embodiments, there is provided a composition comprising: 1-95%phytosterols; 1-95% essential fatty acid complexes; and 1-95%antioxidants. The composition may further include 1-95% filler, forexample, micro-cellulose filler. In other embodiments, 5-95% of thephytosterols may be beta-sitosterol.

A further innovative feature of the invention is the discovery that theessential fatty acid complexes together are not only themselves animmune system modulator, but also facilitates the body's absorption ofbeta-sitosterol and antioxidants.

This invention is a composition of natural products that is designed tostimulate the body to marshal its own natural defenses against allergiesand other immune related disorders, breast, colon and prostate cancer,and AIDS, as opposed to alleviating or preventing these disorders withchemical or synthetic products. Many of the traditional treatments havesevere side effects, which in the case of prostate cancer can includeimpotence and incontinence, and in the case of chemotherapy, severedepression of the immune system. Traditional treatments also only treatthe tumors, i.e. the symptoms, and not the underlying cause, and in thecase of chemotherapy, leave dead tissue to be removed from the body.

One of the suspected causes of cancer is free radical damage on cellularcomponents such as DNA or the cell membranes. Free radicals are thenatural by-products of many processes within and among cells. They arealso created by exposure to various environmental factors such astobacco smoke and radiation. Once formed these highly reactive radicalscan start a chain of reactions that can result in damage to cell walls,certain cell structures, and the genetic material within the cells suchas DNA.

In the preferred form this composition contains one of the world's mostadvanced anti-oxidant complexes, Enzogenol™, produced by EnzoNutraceuticals through a patented process that creates an aqueousextract from Pinus Radiata (pine bark), which is a superior source ofproanthocyanidins and flavonoids. This technically advanced anti-oxidantcomplex provides an excellent source of free radical scavengers:

In addition to scavenging free radicals, the Enzogenol™ in thiscomposition can stimulate phagocytosis, an immune system response inwhich cells ingest and eliminate harmful micro-organisms and foreignheavy metals, and hence facilitates the removal of the by-products ofcell death.

Research has shown that the Human Immune System can be stimulated bybeta-sitosterol. This composition contains this remarkable phytonutrientwhich has been shown to be effective in inhibiting cancer cell growthand accelerating cancer cell death in breast, prostate and colon cancer.It can also reduce metastases to lymph nodes.

Research at the University of New York at Buffalo by Awad, Downey andFink has shown that beta-sitosterol can inhibit breast cancer cellgrowth by 66% after 3 days and 80% after 5 days. It can also increasethe rate of cell death by 600% (Int. J. Mol. Med 2000 May 5(5)).Research by the same group has also shown that beta-sitosterol caninhibit prostate cancer cell growth by 24% and increase cell death by400% (Nutr Cancer 1998; 32 (1)). Similar research on colon cancer cellsshowed a reduction in cell growth of 55% (Anti Cancer Res, 1998March-April 18 (2a)). The exact mechanism behind the inhibition ofcancer cell growth and the increase in apoptosis is not yet fullyunderstood, but may be related to the increase in T-cell proliferationproduced by beta-sitosterol (Int. Jnl. Immuno Pharmacol 1996 vol 18Bouic et al). Beta-sitosterol has been shown to maintain CD4 lymphocytecount in AIDS patients (AIDS Bulletin Summer 97).

The potential benefits of the antioxidants together with thebeta-sitosterol to scavenge free radicals and to kill and remove tumorscan only be realized if the human body actually absorbs both theanti-oxidant complex and the betasitosterol. The role of the essentialfatty acid complex is to ensure that the antioxidant complex and thebeta-sitosterol are absorbed. The essential fatty acid complex itself isa blend of essential fatty acids, amino acids, proline rich peptides,short chain polypeptides and enzymes. This complex also happens to be animmune system modulator, and consequently also supports the body'sattack on cancer by promoting and supporting the T-cell proliferationinitiated by the beta-sitosterol.

The combined effects of the antioxidant complex, the beta-sitosterol andthe essential fatty acid complex on breast, colon and prostate cancersare to reduce the risk of the formation of new cancers through the moreeffective scavenging of free radicals, to inhibit the growth andincrease the rate of cell death of the tumors, and to assist the body indisposing of dead tumor cells.

This formulation has been used to treat a 68 year old male with a PSA of17. Over a six month period his PSA, (the normal measure of prostatecancer which would normally increase over time), dropped to 11.8, areduction of over 30%, with no side effects.

The combined immune modulating properties of beta-sitosterol,antioxidant complex, and the essential fatty acid complex also make thisformulation an excellent anti-allergen with no side effects which isalso new and unique in the field of anti-allergens. This formulation hasbeen used to treat several adults with ages ranging between 22 and 52with excellent results occurring in the space of five to six days, againwith none of the side effects normally associated with anti-allergens.

Early trials of this unique formulation have also shown the capacity toalleviate such immune related disorders as rheumatoid arthritis, againwith none of the adverse side effects normally associated with theremedies traditionally used in this area.

In addition, the effectiveness of beta-sitosterol in maintaining the CD4lymphocyte count in AIDS patients can be increased by the addition ofthe antioxidant and essential fatty acid complexes in this formulation,again providing an important new application for this new and uniqueformulation.

Normally capsules are taken once a day, in a preferred embodiment, atleast one hour or at least half an hour before and at least 2 hoursafter food, with water or fruit juice (NOT milk or any other fatcontaining liquid). As will be appreciated by one of skill the art,these time frames and frequencies are suggested for most users, butother users may find best results taking the capsule more frequently orless frequently, or, for example, with meals or closer to meals.

The term “phytosterols” is to be construed to include, but not to berestricted to, beta-sitosterol, stigmasterol, campesterol,brassicasterol together with their associated glucosides.

The term “antioxidant complex” is to be construed to include, but not tobe restricted to, a broad range of proanthocyanidins, flavonoids andpolyphenols.

The term “essential fatty acid complex” is to be construed to include,but not restricted to, amino acids, essential fatty acids, peptides,proline rich poly peptides, and various digestive enzymes.

The following are examples of usages, showing the utility of thecomposition.

EXAMPLE I Allergies

The composition was used by an individual to treat seasonal allergies.The individual found that one or more of the symptoms associated withallergies, including runny nose, red itchy eyes, aching joints and afeeling of always being tired, were ameliorated following taking thecomposition. Specifically, users found that the symptoms wereameliorated after 24 hours and remained symptom free after 2 weeks bytaking one dosage or capsule per day.

Another individual who suffered from moderate asthma and allergies todust and animal hair found that taking the composition relieved orameliorated symptoms including itchy and watery eyes and a runny nose.This individual also reported increased energy and an improved sense ofwell-being.

Another individual who suffered from allergies to dust mites which ledto a chronically stuffed nose that would drip constantly found thatthese symptoms were ameliorated following taking the composition over aninitial regimen and then taking the composition daily on an as neededbasis.

Another individual reported allergies to pollen and other seasonalallergies and found that taking the composition over a period of a fewweeks ameliorated the allergy-related symptoms, including itchy eyes andsneezing.

EXAMPLE 2 Asthma

An individual who suffered from asthma found that following taking thecomposition regularly for 3 months, the asthma-associated symptoms wereameliorated.

Another individual who suffered from asthma and food allergies and hadsymptoms including itchy eyes and a runny nose found that within 2 weeksof taking the composition on a regular basis, she was off theantihistamines and her stuffy nose was completely gone. Furthermore, shefound that her asthma progressively improved and was able to eat foodsthat she couldn't eat before without having immediate allergicreactions.

EXAMPLE 3 Colds & Flu

One individual who had contracted pneumonia took two capsules of thecomposition per day over a five day period and found that the symptomsassociated with pneumonia were ameliorated.

Another individual who also had a cold develop into pneumonia found thatthe symptoms associated with the illness were ameliorated during a twomonth regimen of taking the composition daily.

Another individual took the composition regularly to prevent colds or“flu”. This individual found less frequent occurrences of colds and“flu” and also found that taking the composition on a regular basisprovided “an all over feeling of wellbeing”.

EXAMPLE 4 Diabetes

A diabetic who was using insulin 3 times per day for the last 12 years,at an average of 50 units a day pf insulin, consisting of 20 units inthe morning, 20 units in the afternoon and 10 units before bed foundthat after having taken the composition daily for 3 weeks that theindividual was able to eliminate the night-time units.

EXAMPLE 5 Eczema

An individual who suffered from eczema, particularly under her armpitson both sides, would also get eczema on her hands whenever her handswere exposed to any chemicals or detergents (ie doing any cleaning) andeven after folding the laundry. She managed the eczema with cortisonecream-a few times per week. After 2 months of taking the compositionalong with eliminating certain food allergies, her eczema was 90% gone(in places where it had been chronic for years) and her hands (whichused to be effected almost constantly) were completely clear.

EXAMPLE 6 Fibromyalgia

An individual who suffered from fibromyalgia found that taking one doseof the composition one hour before breakfast helped ameliorate symptomsincluding pain and insomnia.

Another individual with fibromyalgia reported wakening 34 times pernight with severe cramps in their feet and calves. Following taking thecomposition on a regular dosage regimen, this individual foundamelioration of many of the symptoms associated with the fibromyalgia.

EXAMPLE 7 Hepatitis C

An individual who suffers from hepatitis C reported a progressiveincrease in energy during the first 2 weeks of taking the composition.

EXAMPLE 8 HIV

An individual having an HIV infection reported that their CD4 levelclimbed by 300 points following a 3 month regimen of taking thecomposition.

EXAMPLE 9 Multiple Sclerosis

An individual with multiple sclerosis reported increased energy andimproved balance on taking the composition.

EXAMPLE 10 Urinary Flow

An individual suffering from prostate enlargement reported thatfollowing taking the composition, the time between urinations wasincreased and also reported increased flow.

EXAMPLE 11 Pelvic Inflammatory Disease

Several users have reported that the composition as described abovealleviated one or more of the symptoms associated with pelvicinflammatory disease.

EXAMPLE 12 Rheumatoid Arthritis

An individual with rheumatoid arthritis reported that following takingtwo capsules of the composition per day for two weeks, many of thesymptoms associated with the rheumatoid arthritis were ameliorated.

Dosage and Administration

The recommended adult dosing regimen is currently one capsule per day byoral ingestion. In most embodiments, the supplement is taken with wateror juice (not milk), on an empty stomach 45 minutes before eating. Onecapsule a day normally maintains general health. More serious conditionsmay require two capsules per day, one in the morning and one at night.The dosing regimen can be changed, for example, to two capsules per dayfor the first week, followed by one capsule per day.

Pharmacology

Phytosterols:

Beta-sitosterol is not synthesized endogenously in mammals. Animalstudies have demonstrated its intestinal absorption in mammals isminimal, possibly as little as 5% of total dietary beta-sitosterolconsumed. In contrast, intestinal absorption of cholesterol is 45-54% ofintake. Unlike cholesterol, beta-sitosterol is secreted into the bileand is esterified outside the intestinal wall at a much slower rate.After secretion into the bile, beta-sitosterol is stored in thegallbladder, then released intermittently into the duodenum, andsubsequently incorporated into feces.

It has been proposed that phytosterols inhibit the uptake of dietary andendogenously produced cholesterol from the gut, causing a decrease inserum cholesterol levels (Nguyen 1999). One theory suggests thatcholesterol in the intestine, already marginally soluble, isprecipitated into a non-absorbable state by the presence of addedphytosterols (Hicks and Moreau 2001). A second theory proposes thatcholesterol must enter bile salt and phospholipid containing “mixedmicelles” to be absorbed into the bloodstream. Cholesterol is onlymarginally soluble in these micelles and is displaced by phytosterols,preventing its absorption (Hicks and Moreau 2001). Due to the limitedcapacity in the micelles for carrying cholesterol, compounds withsimilar structures, such as plant sterols, can compete with cholesterolfor this space. Therefore, increasing the amounts of plant sterols mayresult in less cholesterol in mixed micelles and hence, decreasedabsorption of cholesterol from the gut.

Orally ingested flavonoids are largely present as aglycons in theintestine and become absorbed with micelles of bile acids into theepithelium and then into the blood. Through the portal vein, the majorpart of the flavonoids would be delivered to the liver, which decomposesthem (Havsteen 1983, Biochem Pharmacol 32: 1141-1148).

Both EPA and DHA have been found to alter plasma membrane composition,cell-signaling mechanisms, eicosanoid responses, eytokine release, andimmune cell responses (Arslan et al, 2002, Lipids 37: 935-941). Onemechanism for these effects may be that EPA and DHA are incorporatedinto membrane phospholipids by replacing arachidonic acid (AA, n-6 fattyacid). AA is the substrate for the synthesis of eicosanoids, such asfluomboxane A2 and prostaglandin E2 (O'Morain et al. 1990, Scand JGastroenterol 31: 267-272). N-3 fatty acids have a greater affinity forthe cyclo- and lipoxygenase enzymes than n-6 fatty acids and theycompetitively inhibit the formation of prostaglandins and leukotrienes(Drevon 1992). Therefore, increased dietary intake of n-3 fatty acidsmay shift the balance of the eicosanoid production to a lessinflammatory profile (Arslan et al. 2002). Another mechanism for theeffect of n-3 fatty acids is associated with the change in fluidity byincorporation of fatty acids in the cell membranes and an influence onthe activities of membrane-associated enzymes or receptors (Vognild etal. 1998, Lipids 33: 427-436).

This study was designed to investigate whether supplementation with theabove-described supplement comprising plant sterols, antioxidants andessential fatty acids could, in subjects with non-food allergies:

a) Decrease immunological response to allergens

b) Beneficially modify blood lipid parameters

Subjects

Twenty subjects consisting of 16 females and 4 males, aged from 20-50,and who fit specific inclusion/exclusion criteria were recruited. Theinclusion/exclusion criteria were as follows:

Inclusion Criteria:

-   -   Male and female subjects aged 18 years or older    -   For females, confirmation (via pregnancy test) of non-pregnancy        at baseline, and use of acceptable birth control method in        females of childbearing potential    -   No food allergies    -   Willing and able to give informed written consent        Exclusion Criteria:    -   Pregnancy/breast feeding    -   History of clinical significant and unstable cardiovascular,        pulmonary, renal, neurological dermatological, hepatic or        endocrine disease in the past 6 months    -   Change in medication 4 weeks before entry into study    -   History of frequent respiratory infections    -   Diabetes or immune disorder such as lupus erythematosis or        HIV/AIDS    -   Known allergy to the supplement or any of its components    -   History of drug, alcohol or substance abuse in the past 6 months    -   Participation in any clinical trial within 6 weeks preceding day        1 of the study

All participants were recruited from the University of Guelph or theCity of Guelph. At the study screening, participants were given a letterof information and the consent form to sign. Respondents received averbal briefing of the study protocol and the same information inwriting. They signed an informed consent and completed a questionnaireto ensure compliance with the inclusion/exclusion criteria.

All participants were questioned on the nature of their allergies.Eligible subjects were required to have non-food allergies, but wererequired to have allergies to dust, animal dander, animal saliva, molds,etc. In addition, the study required that all subjects had received anallergy test by their physician to verify allergen response, or had beenprescribed medication in the past to treat their allergies.

Experimental Design

This study was conducted as a double blind, placebo-controlled clinicaltrial. Subjects in the treatment group consumed two capsules of theabove-described supplement for the first seven days in order to reachphytosterol threshold levels, followed by a maintenance dosage of 1capsule per day. In these experiments, each capsule contained 300 mg ofphytosterols (117 mg of β-Sitosterol) derived from soy, 20 mg ofEnzogenol™ (antioxidants) extracted from pine bark, and 50 mg ofCellasate™ (a mixture of proteins and essential fatty acids) from seedsand fish oils, although as will be appreciated by one of skill in theart, other suitable formulations as discussed above may also be used.These include embodiments wherein the composition or supplementcomprises: 10-90% phytosterols, 5-85% essential fatty acid complexes,and 5 to 85% antioxidant; or 29% to 75% phytosterols; 10% to 50%essential fatty acid complexes; and 5% to 50% antioxidants as well asembodiments wherein the supplement or composition comprises 29% to 75%beta-sitosterol; 10% to 50% essential fatty acid complexes; and 5% to50% antioxidants as well as other embodiments described herein.

The placebo consisted of a rice flour mixture, identical in appearanceto the supplement, and the consumption pattern for the placebo was thesame for the placebo group as for the treatment group. The supplementand placebo supplementation phase lasted 28 days.

Subjects were required to visit the HNRU human clinical testing unit onthree separate occasions. Subjects came in for baseline testing (day 0)and returned on day 7 of the study and again on day 28, the last day ofthe study. On the initial visit, height, weight and a blood sample weretaken. Blood was analyzed for complete blood cell count (CHC), plasmaDHEA, Cortisol, total HDL and LDL-cholesterol concentrations, andtriglycerides. A Quantikine high sensitivity Elisa kit was used in orderto measure human IL-6 in the plasma collected. Throughout the study,subjects were required to complete a daily journal listing specificallergy symptoms, non-allergy symptoms and any medications taken duringthe supplementation phase.

Results

Immune Parameters

Basophils

The effects of the supplement on immune parameters are presented inTable 1. A number of studies support the belief that human basophilsplay an important role in allergic inflammation. Mast cells andbasophils express the high affinity receptor for IgE (FcepsilonRI) andplay a central role for IgE-associated immediate hypersensitivityreactions and allergic disorders. During allergic reactions, basophilsmigrate from the blood compartment to inflammatory sites, where they actas effector cells in concert with eosinophils. Basophils releasehistamine during inflammation and allergic reactions.

The participants in the treatment group, when compared to the controlgroup, showed a significant reduction in basophil count, while thereduction seen in the control group was non-significant. A reduction inbasophil count may indicate a reduction in histamine release.

Interleukin-6

The immune system also responds to stressors by causing certain immunecells to secrete the pro-inflammatory cytokines, Interleukin-1 (IL-1)and Interleukin-6 (IL-6). These cytokines are both involved ininflammation and IL-6 in particular is thought to worsen the symptoms ofautoimmune diseases and fibromyalgia. Interleukin-6 has been found toact as a growth factor in several tumors and some viruses also use IL-6to replicate. Interleukin-6 also causes calcium to be released frombone, promoting osteoporosis. We must control the release of thesecytokines if we want to enhance immunity and reduce degenerativediseases.

It was noted in the pilot trial that the pro-inflammatory cytokine IL-6levels showed a substantial reduction in the treated group when comparedto the control group.

The supplement has demonstrated that it has an effect onhistamine-containing basophil counts and a reduction of IL-6 levels, andconsequently may substantially alleviate symptoms associated withairborne allergens, asthma and allergic rhinitis.

Cortisol/DHEA Levels

The body has developed mechanisms to protect it from the damagingeffects of stress. The “fight-or-flight” response is one way the bodydeals with extreme situations of stress. Upon realizing we are indanger, the brain sounds an alarm, telling our adrenal glands to secreteadrenaline and cortisol, which mobilizes the body to fight or run. Thisresponse is supposed to be a short-lived reaction yet today most of usare in and out of this state continually. As a result, our immune systembecomes imbalanced, sending out too many inflammatory cytokines. Ouradrenal glands become exhausted, weakening several body systems,especially the cardiovascular and endocrine systems.

DHEA is an abbreviation for dehydroepiandrosterone. It is a hormone madeprimarily by the adrenal or stress glands. Hormones are messengermolecules that influence the function of cells and tissues all over thebody. DHEA and cortisol are the body's long-acting stress hormones andare antagonistic to each other to some degree. Whereas DHEA has ananabolic or building influence, cortisol has a catabolic or tearing downeffect on the body. Both of these effects are essential and these twohormones must be in proper balance for optimal health. How do thesehormones become imbalanced?

By stress maladaptation. Stress maladaptation is the body'sinappropriate response to prolonged stress. The normal reaction of thebody to stress is to produce greater quantities of both cortisol andDHEA. When the stress is gone, the body reduces its output of cortisoland DHEA to resting levels and everything is fine. This is what happenswith short episodes of stress. However, when the stress is prolonged,the body prefers to make increasingly greater amounts of cortisol andless DHEA. How long does it take for this to occur?

One study showed that after just 28 days of continuous stress cortisollevels had climbed to 240 percent of starting values and DHEA haddropped to 15 percent of initial levels. What's even worse is that evenafter the stress is removed, the body sometimes does not recover andbring these hormones back to normal levels, but instead, remains in thestress response mode with high cortisol and low DHEA output. Theconsequences of elevated cortisol and reduced DHEA levels aredevastating: the immune system is compromised with increased risk toinfections, certain cancers, allergies and autoimmune diseases.

A tremendous body of research has shown that when cortisol goes up, DHEAdrops and when DHEA is normal, cortisol also normalizes. Low DHEA levelsare seen in those that are immune compromised, have arteriosclerosis(hardening of the arteries), diabetes and lupus.

Cortisol helps the body maintain homeostasis in the face of stressors,counteracts inflammatory and allergic reactions and controls themetabolism of protein and carbohydrates. Cortisol is a verymisunderstood hormone. Balance is the key. In naturally low doses, itstimulates the immune system and in high doses, as prescribed insynthetic drug form, it can be immune suppressing. Remember thatcortisol plays a role in counteracting inflammatory responses in theimmune system and when cortisol is not available because the adrenalglands have become exhausted from too much stress, inflammation isallowed to continue unchecked. Conversely, too much cortisol and youhave immune suppression.

In the conventional standard of care, any cortisol level within a verybroad range is considered normal, and anything outside that rangeindicates disease. Cortisol production has an ACTH dependent circadianrhythm with peak levels in the early morning and a nadir at night(salivary cortisol ranges can vary from 8.0 to 1.0 in the morning and1.0 to 0.1 in the evening) The factor controlling this rhythm is notcompletely defined and can be disrupted by a number of physical andpsychological conditions. ACTH and cortisol are secreted independent ofcircadian rhythm in response to physical and psychological stress.

In the early stages of adrenal stress, cortisol levels will be too highduring the day and continue rising in the evening. This is called“hyperadrenia”. In the middle stages, cortisol may rise and fallunevenly as the body struggles to balance itself despite the disruptionsof caffeine, carbs and other factors, but levels are not normal and aretypically too high at night. In advanced stages, when the adrenals areexhausted from overwork, cortisol will never reach normal levels(“hypoadrenia”).

None of the participants in the trial were known to have any of theautoimmune diseases that are associated with elevated cortisol levels.The change in cortisol levels noted in the pilot trial appear to be inthe normal range, and neither DHEA nor cortisol levels, nor the ratio ofthese two parameters, showed significant changes at the pa0.05 level.

Cardiovascular Parameters

The effects of the supplement on lipid and lipoprotein parameters areillustrated in Table 2 and Table 3.

Cholesterol is an extremely important biological molecule that has rolesin membrane structure as well as being a precursor for the synthesis ofthe steroid hormones and bile acids. Both dietary cholesterol and thatsynthesized de novo are transported through the circulation inlipoprotein particles. The same is true of cholesterol esters, the formin which cholesterol is stored in cells.

The synthesis and utilization of cholesterol must be tightly regulatedin order to prevent over-accumulation and abnormal deposition within thebody. Of particular importance clinically is the abnormal deposition ofcholesterol and cholesterol-rich lipoproteins in the coronary arteries.Such deposition, eventually leading to atherosclerosis, is the leadingcontributory factor in coronary artery disease.

Cholesterol is minimally soluble in water; it cannot dissolve and travelin the water-based blood stream. Instead, it is transported in the bloodstream by lipoproteins, i.e. protein “molecular-suitcases” which arewater soluble and carry cholesterol and fats internally. The proteinsform part of the surface of the given lipoprotein particle and determinefrom what cells cholesterol will be removed and where it will besupplied.

The largest lipoproteins, which primarily transport fats from theintestinal mucosa to the liver, are called chylomicrons. They carrymostly triglyceride fats and cholesterol (both from food and especiallyinternal cholesterol secreted by the liver into the bile). In the liver,chylomicron particles give up triglycerides and some cholesterol and areconverted into low-density lipoprotein (LDL) particles which carrytriglycerides and cholesterol on to other body cells. In healthyindividuals the low-density lipoprotein (LDL) particles are large andrelatively few in number. Conversely, high numbers of small low-densitylipoprotein (LDL) particles are strongly associated with promotingatheromatous disease within the arteries.

High-density lipoprotein. (HDL) particles transport cholesterol back tothe liver for excretion, but vary considerably in their effectivenessfor doing this. Having large numbers of large HDL particles correlateswith better health outcomes. Conversely, having small amounts of largeHDL particles is strongly associated with atheromatous diseaseprogression within the arteries.

The cholesterol in LDL cholesterol and the cholesterol in HDLcholesterol are identical. The only difference between the two is thecarrier protein molecules (i.e. the lipoprotein).

High cholesterol levels has been shown in many trials to be the sourceof cardiovacular disease. High cholesterol is the best known of all themany threats to a healthy heart. When excess amounts of this waxy,fat-like substance build up along the walls of the arteries, you face adramatically higher risk of a complete blockage, leading to a heartattack or stroke.

At normal levels, cholesterol is not a bad thing. On the contrary, it'san essential raw material used by the body to build cell walls andproduce hormones such as estrogen and testosterone. The body producesits own supply of cholesterol in the liver, and it's found naturally inall animal products (such as meats, eggs, milk, and cheese). It poses aproblem only when the body is unable to use or eliminate excessivesupplies.

As one of a variety of fatty substances in the body, cholesterol isclassified as a lipid. It is carried through the bloodstream attached toproteins, forming complexes called lipoproteins. There are two majortypes of lipoproteins: the low-density lipoproteins (LDL) commonly knownas “bad” cholesterol, and the high-density lipoproteins (HDL) usuallydubbed “good” cholesterol. It's the “bad” LDL cholesterol that tends toform deposits on the artery walls. HDLs, on the other hand, help toclear excess cholesterol from the bloodstream. The ideal situation toaim for then, is a low level of LDL cholesterol, a high level of HDLcholesterol, and a moderate total of both.

The specific objective of this portion of the trial was to determine theeffects of the supplement on blood lipid parameters. Significantreduction was noted in the overall LDL levels of the treatment groupfrom day 0 to day 28. Perhaps what is more interesting is the increasein HDL levels, compared with a relative decrease in the placebo group.

However it is the ratios of various lipids and lipid proteins ratherthan the absolute values that are important in assessing cardiovascularrisk, and consequently these ratios were calculated and tabulated.

A significant decrease in the ratio of TC/HDL, and in the ratio ofLDL/HDL cholesterol in the supplement group was noted. A decrease inthese ratios corresponds to an associated decrease in the risk ofcardiovascular disease (CVD). These ratios are markers for a reductionin the risk of developing atherosclerosis. Consequently these resultsindicate that the supplement is very beneficial to for example thehealth of hypercholesterolemic individuals at risk of developing CVD.

As discussed above, there is provided a method of modifying blood lipidparameters comprising administering to an individual in need of suchtreatment an effective amount of the composition or supplement asdescribed herein. As will be apparent to one of skill in the art, aneffective amount of the supplement refers to the quantity thereofnecessary to reduce the TC/HDL ratio or reduce the LDL/HDL ratio of anindividual in need or desirous of such treatment and such dosages andregimens are enabled herein.

Specifically, as can be seen in table 2, the blood lipid parameters ofthose in the supplement group were modified in that total cholesterolwas reduced by 5.3%, low density lipoprotein was reduced by 15% and HDLwas increased by 4.3%. This is in stark contrast with the resultsobserved for the control group fed the placebo, also shown in Table 2,wherein total cholesterol actually increased by 8.2%, LDL increased by0.7% and HDL decreased by 4.7%.

As summarized on Table 3, the test group administered the supplement orcomposition showed a TC/HDL ratio reduced by 9.4% whereas the control orplacebo group had an increased TC/HDL ratio of 1.7%. Similarly, thesupplement group showed an LDL/HDL ratio decrease of 17.7% whereas theplacebo or control group had only a 0.5% decrease. Finally, thesupplement group showed a total glycerides to HDL increase of 3.0%whereas the control group showed a 20% increase.

Thus, administration of the supplement as described herein has beenshown to reduce total cholesterol, increase HDL, decrease LDL, reducethe TC/HDL ratio and decrease the LDL/HDL ratio in test individualscompared to individuals administered a placebo or control. As such, thesupplement can be used to reduce total cholesterol, increase HDL,decrease LDL, reduce the TC/HDL ratio and decrease the LDL/HDL ratio.

The supplement has an effect on immune parameters and, in particular, onbasophil and possibly IL-6 levels. Given these changes, the supplementhas the potential to substantially alleviate allergic responses.

The supplement could also have an effect in autoimmune diseases such asCrohn's disease or rheumatoid arthritis, or in the ability of subjectsto resist the common cold virus.

This study verified that the supplement is effective in reducingcirculating levels of LDL-cholesterol and increasing circulating levelsof HDL cholesterol. It is of interest to note that there was asignificant decrease in the ratio of TC/HDL, and in the ratio of LDL/HDLcholesterol, in the supplement group. A decrease in these ratioscorresponds to an associated decrease in cardiovascular disease (CVD)risk, because these ratios are markers for a reduction in the risk ofdeveloping atherosclerosis.

While the preferred embodiments of the invention have been describedabove, it will be recognized and understood that various modificationsmay be made therein, and the appended claims are intended to cover allsuch modifications which may fall within the spirit and scope of theinvention.

REFERENCES

-   International Journal of Immuno Pharmacology 1996 Vol 18 Bouic et    al. Beta-sitosterol and beta-sitosterol glucoside, stimulate human    peripheral blood lymphocyte proliferation; implications for their    use as an immunomodulatory vitamin combination.-   International Journal Molecular Medicine 2000 Vol 5 Awad et al.    Inhibition of growth, and stimulation of apoptosis, by    beta-sitosterol treatment of human breast cancer cells.-   Anti Cancer Research 1998 Vol 18 (1 a) Awad, Holtz et al.    Beta-sitosterol inhibits growth of HT-29 human colon cancer cells.-   Nutrition and Cancer 1998 Vol 32 (1) von Holtz, Fink et al.    Beta-sitosterol induces apoptosis in LNCaP human prostate cancer    cells.-   AIDS Bulletin 1997 Vol 6 Bouic. Immodulation in HIV/AIDS: The    Tygerberg/Stellenbosch University Experience.-   South African Journal of Science, 1997 Vol 93 Pegel. The Importance    of Sitosterol and Sitosterolin in human and animal nutrition.-   British Journal of Urology 1997 Vol 80 Klippel et al. A multi    centric, placebo controlled, double blind clinical trial of    betasitosterol for the treatment of benign prostatic hyperplasia.-   The Lancet 1995 Vol 345 Berges et al. Randomized placebo controlled    double blind clinical trial of beta-sitosterol in patients with    benign prostatic hyperplasia.-   The Arthritis Trust of America Summer 98 Bouic. Sterols/Sterolins    the natural, none toxic immuno-modulators and their role in the    control of rheumatoid arthritis.-   Nutrition Reviews 1992 Vol 50 N0. 7 Block. The data support a role    for antioxidants in reducing cancer risks.-   Methods in Enzymology 1990 Vol 186 Bors, Heller et al. Flavanoids as    anti-oxidants: determination of radical scavenging efficiencies.-   The Lancet 1996 Vol 344 Cerruti. Oxi-radicals and cancer.-   Biochemical Pharmacology 1990 Vol 39 No. 11 DeWhalley, Rankin et al.    Flavanoids inhibit the oxidative modification of low density    lipoproteins by macrophages.-   The Lancet 1994 Vol 344 Grishan. Oxidants and free radicals in    inflammatory bowel disease.-   Phytochemistry 1987 Vol 26 No. 9 Husain, Cillard et al. Hydroxyl    radical scavenging activity of flavanoids.-   The Lancet 1994 Vol 344, Jenner. Oxidative damage in    neuerodagenerative disease.-   Free Radicals in Diagnostic Medicine, D. Armstrong, Plenum Press New    York. Free radical scavenging and antioxidant activity of plant    flavanoids.-   New Zealand Journal of Science 1973 Vol 16 Markam and Porter.    Extractives of pinus radiate bark-phenolic components.-   New Zealand Journal of Science 1974 Vol. 17 Porter. Extractives of    pinus radiate bark-procyanidin constituents.-   Free radical Research 1995 Vol 22 No. 4 Rice-Evens, Miller et al.    The relative antioxidant activities of plant derived polyphenolic    flavanoids.-   Biochemical Pharmacology 1998 Vol 37 No 5 Rbak, Gryglewski.    Flavanoids are scavengers of super oxide anions.

Free Radical Biology in Medicine 1990 Vol 9 Yuting, Rongliang et al.Flavanoids as super oxide scavengers and antioxidants. TABLE 1 Theeffects of supplement on specific immune parameters in experimental andplacebo groups from day 0 to day 28 Immune Supplement Supplement ControlControl Parameter Day 0 Day 28 Day 0 Day 28 IgE 472.00 451.00 1335.001127.00 DHEA 6.44 6.44 4.93 4.77 Cortisol 507.00 584.00 490.00 498.00Cortisol/DHEA 94.06 108.36 160.66 141.44 IL-6 1.261 0.937 1.318 1.179WBC 7.41 7.24 7.28 7.13 Lymphocyte 2.16 2.24 2.56 2.60 Segmented 4.654.39 4.11 3.97 Neutrophil Monocytes 0.33 0.34 0.35 0.31 Eosinophils 0.240.20 0.23 0.20 Basophils 0.23 0.01 0.13 0.04

TABLE 2 The effects of supplement on blood lipid parameters inexperimental and placebo groups from day 0 to day 28 Blood LipidSupplement Supplement Control Control Parameter Day 0 Day 28 Day 0 Day28 Total Cholesterol 4.36 4.13 4.87 4.91 LDL 2.27 1.93 2.85 2.87 HDL1.63 1.70 1.48 1.41 TG 1.00 1.09 1.18 1.38

TABLE 3 The effects of supplement on specific cardiovascular ratios inexperimental and placebo groups from day 0 to day 28 CardiovascularParameter Supplement Supplement Control Control Ratios Day 0 Day 28 Day0 Day 28 TC/HDL 2.88 2.61 3.50 3.56 LDL/HDL 1.58 1.30 2.11 2.10 TG/HDL0.66 0.68 0.85 1.02

1. A method of modifying blood lipid parameters comprising administeringto an individual in need of such treatment an effective amount of acomposition comprising: 10% to 90% phytosterols; 5% to 85% essentialfatty acid complexes; and 5% to 85% antioxidants.
 2. The methodaccording to claim 1 wherein the composition comprises: 29% to 75%phytosterols; 10% to 50% essential fatty acid complexes; and 5% to 50%antioxidants.
 3. The method according to claim 1 wherein the compositioncomprises 29% to 75% beta-sitosterol; 10% to 50% essential fatty acidcomplexes; and 5% to 50% antioxidants.
 4. The method according to claim1 wherein the composition comprises 300 mg phytosterols; 50 mg essentialfatty acid complexes; 20 mg antioxidants; and 30 mg micro-cellulosefiller.
 5. The method according to claim 1 wherein the total cholesterolto high density lipoprotein ratio of the individual is reduced.
 6. Themethod according to claim 1 wherein the low density lipoprotein to highdensity lipoprotein ratio of the individual is reduced.